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Evidence that Porphyromonas (Bacteroides) gingivalis fimbriae function in adhesion to Actinomyces viscosus.

机译:牙龈卟啉单胞菌菌毛在粘着放线菌中起作用的证据。

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摘要

Porphyromonas (Bacteroides) gingivalis adheres to gram-positive bacteria, such as Actinomyces viscosus, when colonizing the tooth surface. However, little is known of the adhesins responsible for this interaction. A series of experiments were performed to determine whether P. gingivalis fimbriae function in its coadhesion with A. viscosus. Fimbriae typical of P. gingivalis were isolated from strain 2561 (ATCC 33277) by the method of Yoshimura et al. (F. Yoshimura, K. Takahashi, Y. Nodasaka, and T. Suzuki, J. Bacteriol. 160:949-957, 1984) in fractions enriched with a 40-kDa subunit, the fimbrillin monomer, P. gingivalis-A. viscosus coaggregation was inhibited by purified rabbit antifimbrial immunoglobulin G (IgG) at dilutions eightfold higher than those of preimmune IgG, providing indirect evidence implicating P. gingivalis fimbriae in coadhesion. Three types of direct binding assays further supported this observation. (i) Mixtures of isolated P. gingivalis fimbriae and A. viscosus WVU627 cells were incubated for 1 h, washed vigorously with phosphate-buffered saline (pH 7.2), and subjected to electrophoresis. Transblots onto nitrocellulose were probed with antifimbrial antiserum. Fimbrillin labeled positively on these blots. No reaction occurred with the control protein, porcine serum albumin, when blots were exposed to anti-porcine serum albumin, (ii) A. viscosus cells incubated with P. gingivalis fimbriae were agglutinated only after the addition of antifimbrial antibodies. (iii) Binding curves generated from an enzyme immunoassay demonstrated concentration-dependent binding of P. gingivalis fimbriae to A. viscosus cells. From these lines of evidence, P. gingivalis fimbriae appear to be capable of binding to A. viscosus and mediating the coadhesion of these species.
机译:当在牙齿表面定植时,牙龈卟啉单胞菌(Bacteroides)粘附于革兰氏阳性细菌,例如粘性放线菌。但是,对于引起这种相互作用的粘附素知之甚少。进行了一系列实验,以确定牙龈卟啉单胞菌在其与粘菌曲霉的共粘附中是否起作用。通过Yoshimura等人的方法从菌株2561(ATCC 33277)中分离出典型的齿龈假单胞菌的菌毛。 (F.Yoshimura,K.Takahashi,Y.Nodasaka,和T.Suzuki,J.Bacteriol.160:949-957,1984),其级分富含40kDa亚单位,即纤颤蛋白单体,P.gingivalis-A。纯化的兔抗纤维化免疫球蛋白G(IgG)可以抑制粘性共聚集,其稀释度比免疫前IgG高八倍,从而间接证明了牙龈卟啉单胞菌菌毛共粘连。三种类型的直接结合测定进一步支持了这一观察。 (i)将分离出的牙龈卟啉单胞菌菌毛和黏稠曲霉WVU627细胞的混合物孵育1小时,用磷酸盐缓冲液(pH 7.2)强烈洗涤,并进行电泳。用抗纤维蛋白抗血清探测到硝酸纤维素上的转运蛋白。在这些印迹上阳性标记的是菌纤蛋白。当印迹暴露于抗猪血清白蛋白时,对照蛋白猪血清白蛋白未发生反应。(ii)仅在添加抗纤维抗体后凝集与牙龈卟啉单胞菌菌丝一起培养的粘液曲霉细胞。 (iii)从酶免疫测定法产生的结合曲线证明了齿龈丙酸杆菌菌毛与粘液曲霉细胞的浓度依赖性结合。从这些证据来看,齿龈假单胞菌似乎能够与粘性粘菌结合并介导这些物种的共粘连。

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    Goulbourne, P A; Ellen, R P;

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  • 年度 1991
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